An analysis of cattle farmers' perceptions of drivers and barriers to on-farm control of Escherichia coli O157.

Structural equation modelling and survey data were used to test determinants' influence on farmers' intentions towards Escherichia coli O157 on-farm control. Results suggest that farmers more likely to show willingness to spend money/time or vaccinate to control Escherichia coli O157 are those: who think farmers are most responsible for control; whose income depends more on opening farms to the public; with stronger disease control attitudes; affected by outbreaks; with better knowledge and more informed; with stronger perceptions of biosecurity measures' practicality; using a health plan; who think farmers are the main beneficiaries of control; and whose farms are dairy rather than beef. The findings might suggest that farmers may implement on-farm controls for E. coli O157 if they identify a clear hazard and if there is greater knowledge of the safety and efficacy of the proposed controls.

Associations between the presence of virulence determinants and the epidemiology and ecology of zoonotic Escherichia coli.

The severity of human infection with pathogenic Escherichia coli depends on two major virulence determinants (eae and stx) that, respectively, produce intimin and Shiga toxin. In cattle, both may enhance colonization, but whether this increases fitness by enhancing cattle-to-cattle transmission in the field is unknown. In E. coli O157, the almost uniform presence of the virulence determinants in cattle isolates prevents comparative analysis. The availability to this study of extensive non-O157 E. coli data, with much greater diversity in carriage of virulence determinants, provides the opportunity to gain insight into their potential impact on transmission. Dynamic models were used to simulate expected prevalence distributions for serogroups O26 and O103. Transmission parameters were estimated by fitting model outputs to prevalence data from Scottish cattle using a Bayesian Markov chain Monte Carlo (MCMC) approach. Despite similar prevalence distributions for O26 and O103, their transmission dynamics were distinct. Serogroup O26 strains appear well adapted to the cattle host. The dynamics are characterized by a basic reproduction ratio (R(0)) of >1 (allowing sustained cattle-to-cattle transmission), a relatively low transmission rate from environmental reservoirs, and substantial association with eae on transmission. The presence of stx(2) was associated with reduced transmission. In contrast, serogroup O103 appears better adapted to the noncattle environment, characterized by an R(0) value of <1 for plausible test sensitivities, a significantly higher transmission rate from noncattle sources than serogroup O26, and an absence of fitness benefits associated with the carriage of eae. Thus, the association of eae with enhanced transmission depends on the E. coli serogroup. Our results suggest that the capacity of E. coli strains to derive fitness benefits from virulence determinants influences the prevalence in the cattle population and the ecology and epidemiology of the host organism.

Emissions of methyl halides and methane from rice paddies.

Methyl halide gases are important sources of atmospheric inorganic halogen compounds, which in turn are central reactants in many stratospheric and tropospheric chemical processes. By observing emissions of methyl chloride, methyl bromide, and methyl iodide from flooded California rice fields, we estimate the impact of rice agriculture on the atmospheric budgets of these gases. Factors influencing methyl halide emissions are stage of rice growth, soil organic content, halide concentrations, and field-water management. Extrapolating our data implies that about 1 percent of atmospheric methyl bromide and 5 percent of methyl iodide arise from rice fields worldwide. Unplanted flooded fields emit as much methyl chloride as planted, flooded rice fields.

Cefixime-tellurite rhamnose MacConkey agar for isolation of Vero cytotoxin-producing Escherichia coli serogroup O26 from Scottish cattle and sheep faeces.

AIMS: To compare rhamnose MacConkey agar supplemented with cefixime and tellurite (CT-RMac) and tryptone bile X-glucuronide (TBX) agars as isolation media for Vero cytotoxin-producing Escherichia coli (VTEC) serogroup O26 from animal faeces. METHODS AND RESULTS: Nine VTEC O26 were isolated from sheep faeces; out of which six were isolated only on CT-RMac and one was isolated only on TBX. One hundred and twelve VTEC O26 were isolated from calf faeces; out of which 97% were from CT-RMac and 52% were from TBX. In a study of E. coli O26 strains, 84% of VT-positive O26 did not ferment rhamnose when compared with 16% of VT-negative O26. VT-positive (19%) and VT-negative (39%) E. coli O26 strains did not grow on CT-RMac agar. CONCLUSIONS: It is important to consider that VTEC O26 strains either may ferment rhamnose or may be sensitive to the CT supplement of CT-RMac agar. SIGNIFICANCE AND IMPACT OF THE STUDY: This work compares CT-RMac and TBX agars as isolation medium for VTEC O26 from Scottish animal faeces and highlights that VTEC O26 may be missed if only CT-RMac agar is used.

Evaluation of a survey approach to estimating the prevalence of cattle carrying antimicrobial-resistant Escherichia coli.

A random survey of farms in the Highlands and Islands of Scotland provides estimated of the prevalence of calves, finishers and cows carrying ampicillin, apramycin and/or nalidixic acid resistant Escherichia coli. While the survey provides information on the geographical variation in risk, the results are of limited value for interpreting causality.

Simultaneous measurement of thyroxine and triiodothyronine peripheral turnover kinetics in man.

Serum triiodothyronine (T(3)) kinetics in man have been difficult to define presumably due to the interference of iodoproteins generated during the peripheral metabolism of T(3). The use, in the present study, of an anion-column chromatographic method for separation of serum T(3) as well as thyroxine (T(4)) from these iodoproteins has overcome this technical handicap. Simultaneous measurement of serum (125)I-T(3) and (131)I-T(4) kinetics were performed in 31 subjects from the clinical categories of euthyroid, primary hypothyroid, thyrotoxic and posttreatment hypothyroid Graves' disease, factitial thyrotoxic, and idiopathically high and low thyroxinebinding globulin states. The normal mean T(3) fractional turnover rate (kT(3)) was 0.68 (half-life = 1.0 days), increased in toxic Graves' disease patients to 1.10 (half-life = 0.63 days), and decreased in primary hypothyroid patients to 0.50 (half-life = 1.38 days). The mean T(3) equilibration time averaged 22 hr except in hypothyroid and high thyroxine-binding globulin (TBG) patients where the equilibration period was delayed by 10 hr. The mean T(3) distribution space in normal subjects was 38.4 liters. This was reduced in subjects with high TBG levels (26 liters) and increased in patients with low TBG and in all hyperthyroid states (53-55 liters). The normal serum T(3) concentration was estimated by radioimmunoassay to be 0.106 mug/100 ml. Combined with the mean T(3) clearance value of 26.1 liters/day, the calculated T(3) production rate was 27.6 mug/day. The mean T(3) production rate increased to 201 mug/day in thyrotoxic Graves' disease patients and was reduced to 7.6 mug/day in primary hypothyroid subjects. T(3) production rate was normal in subjects with altered TBG states. The ratio of T(3) to T(4) production rate in normal subjects was 0.31 and was unchanged in patients with altered TBG values. This ratio was increased in all Graves' disease patients with the highest value being 0.81 in the posttreatment hypothyroid Graves' disease group. This apparent preferential production of T(3) may have been responsible for the retention of rapid turnover kinetics for T(3) and T(4) observed in treated Graves' disease patients. The finding that factitial thyrotoxic patients also displayed similar rapid T(3) and T(4) turnover kinetics indicates that these alterations are not a unique feature of Graves' disease per se. When comparing the peripheral turnover values for T(3) and T(4) in man, it is apparent that alterations in metabolic status and serum TBG concentration influence both hormones in a parallel manner; however, changes in metabolic status seem to have a greater influence on T(3) kinetics while alterations in TBG concentrations have a greater effect on T(4). These observations probably relate to the differences in TBG binding affinity and peripheral tissue distribution of these two hormones.

What is antibiotic resistance and how can we measure it?

Antibiotic resistance is being found with increasing frequency in both pathogenic and commensal bacteria of humans and animals. Quantifying resistance within and between bacterial and host populations presents scientists with complex challenges in terms of laboratory methodologies and sampling design. Here, we discuss, from an epidemiological perspective, how antibiotic resistance can be defined and measured and the limitations of current approaches.

Identification of cattle infected with bovine virus diarrhoea virus using a monoclonal antibody capture ELISA.

A monoclonal antibody capture enzyme linked immunosorbent assay (ELISA) has been developed to detect pestivirus-specific antigen in the leucocytes of cattle infected with bovine virus diarrhoea virus (BVDV). A blind trial was conducted to compare the specificity of the ELISA with conventional tissue culture virus isolation on 215 blood samples submitted for BVDV diagnosis from cattle throughout Scotland. One hundred and sixty seven samples were negative by both ELISA and virus isolation and 47 samples were positive by both tests. One blood was negative by ELISA and positive by virus isolation.

Characterisation of Listeria ivanovii isolates from the UK using pulsed-field gel electrophoresis.

Forty-three Listeria ivanovii isolates were collected in the UK between 1991 and 1997 from: 35 animal infections; two human infections; five foods; and one environmental source. A further two type strains of L. ivanovii (subsp. ivanovii and subsp. londoniensis) were obtained from a culture collection. These bacteria were characterised by conventional phenotypic methods and by pulsed-field gel electrophoresis (PFGE) using ApaI and SmaI. Forty-two of the isolates from the UK were identified as L. ivanovii subsp. ivanovii and the remaining culture as L. ivanovii subsp. londoniensis. Six and four PFGE profiles were obtained using ApaI and SmaI digestion respectively; six composite profiles were obtained combining the results for both enzymes. The PFGE profile of the UK L. ivanovii subsp. londoniensis (isolated from processed shrimps) was similar to the type strain of this subspecies and differed from all of the L. ivanovii subsp. ivanovii tested. The majority of isolates (38 out of 45) belonged to one profile showing that the UK population of this bacterium is much less genetically diverse than similar studies have shown for Listeria monocytogenes.

PCR methods and plasmid profile analysis for characterisation of Histophilus ovis strains.

The value of polymerase chain reaction (PCR)-based DNA fingerprinting and plasmid profile analysis for differentiation of Histophilus ovis isolates was assessed. Nineteen isolates of H. ovis were typed by PCR-ribotyping, repetitive extragenic palindromic element (REP)-PCR and enterobacterial repetitive intergenic consensus (ERIC)-PCR. These methods distinguished five types by PCR-ribotyping, 11 types by REP-PCR and seven types by ERIC-PCR. The ribotyping method produced a relatively simple pattern and a small number of distinct types and was useful for differentiation of H. ovis from the phenotypically similar organism, Haemophilus somnus. REP- and ERIC-PCR both produced complex banding patterns, but increased the discrimination between strains. Plasmids were found in 12 of the 19 isolates and there were four different plasmid profiles. A combination of the PCR methods and plasmid profile analysis provided a high resolution typing method for H. ovis.

Voltage dependency of light-evoked on-off transient amacrine cell responses in carp retina.

The voltage dependency of the ON and the OFF components of transient amacrine cell responses was studied using two-electrode voltage-clamp and current-clamp techniques in the isolated retina of the carp. The two independent approaches gave similar data. When cells were voltage clamped near their resting potentials, both response components were associated with transient inward currents. Hyperpolarization increased response size (current or voltage) whilst depolarization decreased it. Response reversal, or a tendency for it, occurred at membrane potentials significantly more positive than the resting level with some quantitative variability. These data support the view that the ON-OFF depolarizations represent basically excitatory postsynaptic potentials and that the transience of the responses cannot mainly be due to any voltage-dependent conductance.

Binocular fields with lateral-eyed vision.

A simple geometrical model is constructed of the binocular field in animals with laterally directed eyes (such as the pigeon). Height and width are derived as a function of elevation, for eye-centred and for cyclopean co-ordinates. Stereoscopic vision in the frontal field is considered, and it is pointed out that convergence movements can facilitate stereopsis even in an afoveate periphery.

Voltage clamp study of electrophysiologically-identified horizontal cells in carp retina.

Passive membrane properties and electromotive force of light modulated currents of L-, R/G-type and rod-driven horizontal cells were studied by voltage-clamp using double-barrelled micro-electrodes whilst perfusing with 5 microM dopamine to uncouple the gap junctions. Input impedances of horizontal cells in darkness were 31 +/- 1.4 M omega (mean +/- SE, n = 63); the resting potentials were -37 +/- 1.3 mV. Current-voltage relationships had regions of both inward and outward rectification and a region of negative resistance was commonly observed. Reversal potentials of light modulated currents were estimated on average to be -7 +/- 4 mV (n = 14), which is consistent with the involvement of K+ and Na+ and/or Ca2+ gradients. Importantly in R/G cells both depolarizing and hyperpolarizing components of the response had essentially the same reversal potential.

The projection of the visual field upon the retina of the pigeon.

Co-ordinates in the visual field of the pigeon eye were located in the eye cup anatomically by marking the position of the trans-scleral image. The retinal horizon and vertical meridian at azimuths 0 deg (frontal field) and 90 deg (lateral field) were located. Approximately 18 deg of binocular field are available in frontal vision and the frontal meridian projects to temporal retina outside the red field. Spatial distributions of ganglion cells, displaced ganglion cells, and centrifugal terminals were located in the eye cup. Retinal magnification factors in the posterior pole and temporal periphery are close to 120 microns/deg.

Development and evaluation of an indirect ELISA to detect antibodies to abortion strains of Chlamydia psittaci in sheep sera.

A novel indirect enzyme-linked immunosorbent assay (ELISA) for antibodies against abortion strains of Chlamydia psittaci (C. psittaci) has been developed. The antigen used was chlamydial elementary bodies treated sequentially with N-lauroyl sarcosine and n-octyl-beta-D-glucopyranoside and finally solubilized with N-lauroyl sarcosine and dithiothreitol. Treating the antigen with sodium periodate after coating of the plates increased the specificity for antibodies to abortion strains. The test was evaluated initially with sera from experimentally infected sheep and an uninfected control group. These sheep were monitored for lambing performance and infection status. When used in conjunction with the indirect micro-immunofluorescence test (MIF), the ELISA was able to identify as negative all twenty-five sera from ewes that had no typical placental lesions and identified as positive twenty of twenty-one sera from infected ewes that had either typical placental lesions or had been found positive by isolation of chlamydia in cell culture. The combination of ELISA and MIF was also able to discriminate correctly groups of sera from six flocks with a history of infection from four known uninfected flocks.

Photoreceptoral basis for altered dark-adaptation in retinal degeneration (RCS) rats.

The 'fast PIII' rod mediated photoreceptor response to flashes of monochromatic light has been investigated in isolated retinae of the RCS rat at age 30 days. A loss of both sensitivity and maximum response amplitude were found, with a concurrent slowing of the response kinetics. These observations indicate a rod abnormality which may underly the altered adaptational properties in these retinae.

Clinical and serum antibody responses to lambs to infection by Listeria monocytogenes.

Oral dosing of lambs with 1 x 10(10) colony forming units of Listeria monocytogenes daily for three days produced no clinical signs but protected the animals against bacteraemia following subsequent homologous subcutaneous challenge. Following the subcutaneous injections, comparison with controls revealed significantly lower rectal temperatures and a significant difference in positive blood cultures. In both groups signs of systemic illness were unremarkable. However, two and 10 days after the subcutaneous challenges neurological signs developed in two lambs. L monocytogenes was isolated from the brain of one lamb and histopathological lesions of listeric encephalomyelitis were demonstrated in both. After oral infection antibodies to L monocytogenes whole cell antigen were detectable in serum agglutination tests and by ELISA. Serological responses to flagellin were examined by ELISA and to listeriolysin O by immunoblotting. The responses of the animals to flagellin were weak and inconsistent, but antibodies to listeriolysin O were detectable after both oral and subcutaneous challenge. The subclass of antibody involved in this response was shown to be predominantly IgG1.

Pyrolysis mass spectrometry of Listeria monocytogenes isolates from sheep.

Forty-eight isolates of Listeria monocytogenes from sheep and silage, involved in five small outbreaks of listeriosis, were compared by pyrolysis mass spectrometry (PyMS). The method clustered isolates from single animals, and showed that epidemiologically associated isolates were closely related to each other. PyMS is a simple technique capable of analysing large numbers of samples daily, and its application in veterinary studies should help to elucidate the epidemiology of listeriosis.

A review of Listeria monocytogenes and listeriosis.

Following the initial isolation and description in 1926 Listeria monocytogenes has been shown to be of world-wide prevalence and is associated with serious disease in a wide variety of animals, including man. Our knowledge of this bacterial pathogen and the various forms of listeriosis that it causes has until recently been extremely limited, but recent advances in taxonomy, isolation methods, bacterial typing, molecular biology and cell biology have extended our knowledge. It is an exquisitely adaptable environmental bacterium capable of existing both as an animal pathogen and plant saprophyte with a powerful array of regulated virulence factors. Most cases of listeriosis arise from the ingestion of contaminated food and in the UK the disease is particularly common in ruminants fed on silage. Although a number of forms of listeriosis are easily recognized, such as encephalitis, abortion and septicaemia, the epidemiological aspects and pathogenesis of infection in ruminants remain poorly understood. The invasion of peripheral nerve cells and rapid entry into the brain is postulated as a unique characteristic of its virulence, but relevant and practical disease models are still required to investigate this phenomenon. This review offers an up to date introduction to the organism with a description of virulence determinants, typing systems and a detailed account of listeriosis in animals. Experimental and field papers are reviewed and further sections deal with the diagnosis, treatment and control of listeriosis in animals. A final part gives an overview of listeriosis in man.

False-negative morphine-augmented cholescintigraphy in a patient with gangrenous cholecystitis.

Intravenous morphine sulfate is commonly used to shorten study time and has been reported not to lower the specificity of hepatobiliary imaging. Although the false-negative rate is low, caution has to be taken in interpreting morphine-enhanced cholescintigraphy. The report presents a false-negative study in a patient with acute gangrenous cholecystitis.